US6794163B2 - Methods to culture circovirus - Google Patents

Methods to culture circovirus Download PDF

Info

Publication number
US6794163B2
US6794163B2 US10/112,540 US11254002A US6794163B2 US 6794163 B2 US6794163 B2 US 6794163B2 US 11254002 A US11254002 A US 11254002A US 6794163 B2 US6794163 B2 US 6794163B2
Authority
US
United States
Prior art keywords
mammalian
circovirus
adenovirus
porcine
functional protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
US10/112,540
Other languages
English (en)
Other versions
US20020177216A1 (en
Inventor
Qiang Liu
Suresh K. Tikoo
Philip Willson
Lorne A. Babiuk
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Saskatchewan
Original Assignee
University of Saskatchewan
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Saskatchewan filed Critical University of Saskatchewan
Priority to US10/112,540 priority Critical patent/US6794163B2/en
Assigned to UNIVERSITY OF SASKACHEWAN reassignment UNIVERSITY OF SASKACHEWAN ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BABIUK, LORNE A., LIU, QIANG, TIKOO, SURESH K., WILLSON, PHILIP
Publication of US20020177216A1 publication Critical patent/US20020177216A1/en
Assigned to UNIVERSITY OF SASKATCHEWAN reassignment UNIVERSITY OF SASKATCHEWAN CORRECTIVE ASSIGNMENT TO CORRECT THE ASSIGNEES NAME. DOCUMENT PREVIOUSLY RECORDED AT REEL 013114 FRAME 0101. Assignors: BABIUK, LORNE A., LIU, QIANG, TIKOO, SURESH K., WILLSON, PHILIP
Priority to US10/840,879 priority patent/US7172899B2/en
Application granted granted Critical
Publication of US6794163B2 publication Critical patent/US6794163B2/en
Priority to US11/654,869 priority patent/US7560278B2/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/10011—Circoviridae
    • C12N2750/10021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/10011—Circoviridae
    • C12N2750/10041—Use of virus, viral particle or viral elements as a vector
    • C12N2750/10043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/10011—Circoviridae
    • C12N2750/10051—Methods of production or purification of viral material
    • C12N2750/10052—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles

Definitions

  • the present invention relates to the field of circovirus and provides compositions and methods for culturing circovirus, in particular porcine circovirus.
  • the present invention relates to methods for culturing porcine circovirus in mammalian cells expressing a mammalian adenovirus E1 gene function.
  • Circoviridae A family of viruses, named Circoviridae, found in a range of plant and animal species and commonly referred to as circoviruses, are characterized as round, non-enveloped virions with mean diameters from 17 to 23.5 nm containing circular, single-stranded deoxyribonucleic acid (ssDNA).
  • the ssDNA genome of the circoviruses represent the smallest viral DNA replicons known.
  • at least six viruses have been identified as members of the family according to The Sixth Report of the International Committee for the Taxonomy of Viruses (Lukert, P. D. et al. 1995 , The Circoviridae , pp. 166-168. In F. A. Murphy, et al. (eds.) Virus Taxonomy, Sixth Report of the International Committee on Taxonomy of Viruses, Arch. Virol. 10 Suppl.).
  • PCV chicken anemia virus
  • BFDV beak and feather disease virus
  • PCV porcine circovirus
  • pigeon circovirus was originally isolated in porcine kidney cell cultures. PCV replicates in the cell nucleus and produces large intranuclear inclusion bodies. See Murphy et al. (1999 , Circoviridae p. 357-361, Veterinary Virology, 3rd ed. Academic Press, San Diego).
  • PCV type 1 PCV 1
  • PCV 2 PCV type 2
  • PCV1 isolated as a persistent contaminant of the continuous porcine kidney cell line PK-15 (ATCC CCL31), does not cause detectable cytopathic effects in cell culture and fails to produce clinical disease in pigs after experimental infection (see Allan G., 1995 , Vet. Microbiol . 44: 49-64; Tischer, I. et al., 1982 , Nature 295:64-66; and Tischer, I. et al., 1986 , Arch. Virol . 91:271-276).
  • PCV2 in contrast to PCV1, is closely associated with post weaning multisystemic wasting syndrome (PMWS) in weanling pigs (see Allan G. et al., 1998 , Europe. J. Vet. Diagn.
  • PCV1 The nucleotide sequences for PCV1 are disclosed in Mankertz, A., et al. (1997 , J. Virol . 71:2562-2566) and Meehan, B. M., et al. (1997 , J. Gen. Virol . 78:221-227) and the nucleotide sequences for PCV2 are disclosed in Hamel, A. L. et al. (1998 , J. Virol .
  • the present invention provides methods for culturing mammalian circovirus comprising: a) obtaining mammalian cells expressing a mammalian adenovirus E1 function, wherein said cells are permissive for mammalian circovirus replication; b) introducing said mammalian circovirus genome, or a portion thereof capable of replication, into said mammalian cells; and c) culturing said mammalian cells under conditions suitable for replication of said mammalian circovirus.
  • the method further comprises recovering said circovirus from said cultured cells.
  • the mammalian circovirus is porcine circovirus, such as for example, porcine circovirus 1 (PCV1) or porcine circovirus 2 (PCV2).
  • the porcine circovirus comprises a chimeric nucleotide sequence.
  • the mammalian cells are of porcine origin.
  • the mammalian cells are porcine retina cells.
  • the mammalian adenovirus E1 function is human adenovirus E1 function. In yet other embodiments, the mammalian adenovirus E1 function is porcine adenovirus E1 function. In further embodiments, the E1 function is E1A and/or E1B function. In yet further embodiments, the mammalian cell expressing the mammalian E1 function is stably transformed with mammalian E1 gene sequences. In other embodiments, the mammalian E1 gene sequence is heterologous to said mammalian cell.
  • the present invention also provides recombinant mammalian cells that express a mammalian adenovirus E1 function and comprise a mammalian circovirus genome, or a portion thereof capable of replication, and wherein said cells are permissive for the replication of said mammalian circovirus.
  • the mammalian circovirus is porcine circovirus, such as for example, porcine circovirus 1 (PCV1) or porcine circovirus 2 (PCV2).
  • the porcine circovirus comprises a chimeric nucleotide sequence.
  • the adenovirus E1 function is human adenovirus E1 function.
  • the E1 function is porcine adenovirus E1 function.
  • the mammalian cell is of porcine origin.
  • the mammalian cell is a porcine retinal cell.
  • the mammalian cell expressing the mammalian E1 function is stably transformed with mammalian adenovirus E1 gene sequences.
  • the mammalian E1 gene sequence is heterologous to said mammalian cell.
  • the present invention also provides methods of preparing a recombinant mammalian cell expressing a mammalian adenovirus E1 function and comprising a mammalian circovirus genome comprising the steps of, a) obtaining a mammalian cell expressing a mammalian adenovirus E1 function; and b) introducing said mammalian circovirus genome, or a portion thereof capable of replication, into said mammalian cell.
  • the method comprises the additional step of culturing the recombinant mammalian cell under conditions suitable for the replication of said mammalian circovirus.
  • the method comprises recovering said circovirus from said cultured cells.
  • the mammalian circovirus is porcine circovirus, such as for example, porcine circovirus 1 (PCV1) or porcine circovirus 2 (PCV2).
  • the porcine circovirus comprises a chimeric nucleotide sequence.
  • the mammalian cells are of porcine origin.
  • the mammalian cells are porcine retina cells.
  • the adenovirus E1 function is human adenovirus E1 function or porcine adenovirus E1 function.
  • the mammalian cell expressing the mammalian adenovirus E1 function is stably transformed with mammalian adenovirus E1 gene sequences.
  • the mammalian E1 gene sequence is heterologous to said mammalian cell.
  • FIGS. 1A-1B provide the characterization and titration of PCV2 virus generated by DNA transfection and extraction from infected VIDO R1 cells by Hirt's method.
  • A PCR using PCV2-specific primers and DNA from PCV2-infected (lane 1) and mock-infected (lane 2) cells.
  • a plasmid containing PCV2 genome was used as a control (lane 3).
  • the 1-kb DNA ladder from GIBCO BRL was loaded in lane M.
  • FIGS. 2A-2B depict titration of PCV2 by immunoperoxidase staining.
  • mock-(A) or PCV2-(B) infected VIDO R1 cells were incubated with rabbit anti-ORF2 polyclonal antibody and biotinylated secondary antibody.
  • the monolayer was developed by diaminobenzidine tetrahydrochloride (DAB).
  • DAB diaminobenzidine tetrahydrochloride
  • FIGS. 3A-3C show the nucleotide sequence (A) (SEQ ID NO:1) and amino acid sequence for ORF 1 (B) (SEQ ID NO:2) and ORF 2(C) (SEQ ID NO:3) of porcine circovirus 2 (PCV2) as described in Genbank accession number AF086834.
  • the present invention relates to compositions and methods for culturing mammalian circovirus, in particular porcine circovirus.
  • the present invention is based on the finding that a porcine cell expressing human E1 function was able to be transfected with a PCV2 virus genome and generated PCV2 virus with a high virus titer.
  • VIDO R1 cell line deposited with the ATCC and having ATCC accession number PTA-155, is a porcine retina cell line transformed with human adenovirus-5 (HAV5) E1, that has been shown to induce the S phase of the cell cycle and transactivate transcription. See, Shenk, T. (1996). “ Adenoviridae : the viruses and their replication” In Fields Virology .
  • VIDO R1 cells were transfected with a PCV2 genome and generated virus at 2 ⁇ 10 7 IU/ml.
  • Circoviridae a family of viruses having round, non-enveloped virions with mean diameters from 17 to 23.5 nm containing circular, single-stranded DNA (ssDNA), are described in The Sixth Report of the International Committee for the Taxonomy of Viruses, supra. Members of the group include the porcine circoviruses, PCV1 and PCV2. Some of the PCVs are known to be pathogenic, such as PCV2, associated with PMWS.
  • Nucleotide sequences for PCV1 are provided in Mankertz, A., et al., 1997 , J. Virol . 71:2562-2566 and Meehan, B. M. et al., 1997 , J. Gen. Virol . 78:221-227.
  • Nucleotide sequences for PCV2 are provided in Hamel, A. L. et al., 1998 , J. Virol . 72:5262-5267; Mankertz, A. et al., 2000 , Virus Res . 66:65-77 and Meehan, B. M. et al., 1998 , J. Gen. Virol . 79:2171-2179.
  • PCV2 Representative strains of PCV2 have been deposited with the European Collection of Cell Cultures, Centre for Applied Microbiology & Research, Porton Down, Salisbury, Wiltshire SP4 OJG, United Kingdom and include accession No. V97100219; accession No. V9700218; accession No. V97100217; accession No. V98011608; and accession No. V98011609. WO 00/77216 also discloses PCV2.
  • PCV2 nucleotide sequences have also been published in Hamel et al., (1998), J. Virol . vol. 72, 6:5262-5267 (GenBank AF027217) and in Morozov et al., (1998), J. Clinical Microb . vol.
  • the present invention encompasses methods of culturing mammalian circovirus and in particular, porcine circovirus (PCV).
  • PCV porcine circovirus
  • the present invention encompasses methods of culturing PCV comprising the PCV nucleotide sequences disclosed herein or known in the art, or ORFs thereof, or portions thereof that are capable of replication.
  • the present invention also encompasses methods of culturing PCV having PCV nucleotide sequences differing through the degeneracy of the genetic code to those disclosed herein or known in the art, or ORFs thereof, or portions thereof capable of replication.
  • the present invention further encompasses methods of culturing PCV comprising PCV nucleotide sequence variations which do not change the functionality or strain specificity of the nucleotide sequence, or ORFs thereof, or portions thereof capable of replication.
  • the present invention also encompasses methods of culturing PCV comprising PCV nucleotide sequences capable of hybridizing to those sequences disclosed herein under conditions of intermediate to high stringency, and methods of culturing PCV comprising mutations of the PCV nucleotide sequence disclosed herein or known in the art, such as deletions or point mutations, or ORFs thereof, or portions thereof, capable of replication.
  • the present invention also encompasses methods of culturing PCV comprising heterologous nucleotide sequences.
  • the present invention encompasses methods of culturing PCV that comprise chimeric circovirus nucleotide sequences, such as, for example, nucleotide sequences from porcine circovirus in fusion with nucleotide sequences from other pathogenic viruses, such as a pathogenic porcine virus, including parvovirus.
  • chimeric circovirus nucleotide sequences such as, for example, nucleotide sequences from porcine circovirus in fusion with nucleotide sequences from other pathogenic viruses, such as a pathogenic porcine virus, including parvovirus.
  • a heterologous nucleotide sequence is one which is not normally associated with the circovirus sequences as part of the circovirus genome or one which is not normally associated with the mammalian cell, respectively.
  • Heterologous nucleotide sequences include synthetic sequences.
  • Hybridization reactions can be performed under conditions of different “stringency”. Conditions that increase stringency of a hybridization reaction are widely known and published in the art. See, for example, Sambrook et al. (1989) at page 7.52. Examples of relevant conditions include (in order of increasing stringency): incubation temperatures of 25° C., 37° C., 50° C.
  • buffer concentrations of 10 ⁇ SSC, 6 ⁇ SSC, 1 ⁇ SSC, 0.1 ⁇ SSC (where SSC is 0.15 M NaCl and 15 mM citrate buffer) and their equivalents using other buffer systems; formamide concentrations of 0%, 25%, 50%, and 75%; incubation times from 5 minutes to 24 hours; 1, 2, or more washing steps; wash incubation times of 1, 2, or 15 minutes; and wash solutions of 6 ⁇ SSC, 1 ⁇ SSC, 0.1 ⁇ SSC, or deionized water.
  • An exemplary set of stringent hybridization conditions is 68° C. and 0.1 ⁇ SSC.
  • the PCV genomes encode several polypeptide sequences, ranging in approximate size from 8 to 35 kD. It is deemed routine to determine open reading frames (ORFs) for porcine circoviruses using standard software such as for example, MacVector® (Oxford Molecular Group Inc., MD 21030). The largest ORF, ORF1, of the two types of PCV shows only minor variation with an identity of 85% (as measured by the clustal program) and has been demonstrated to be the Rep protein in PCV1 (Mankertz, A., et al., 1998 , J. Gen. Virol . 79:381-384).
  • PCV2 ORF2 has been identified as a major structural protein that can form viral capsid-like particles in insect cells infected with ORF2 expressing recombinant baculovirus. See Nawagitgul, P. et al., 2000 , J. Gen. Virol . 81:2281-2287.
  • a recombinant vector comprising a PCV genome or an ORF thereof, or a portion thereof, such as an antigenic region, is constructed by in vitro recombination between a plasmid and a PCV genome.
  • the PCV genome is a PCV2 genome.
  • a recombinant vector comprising a PCV genome or an ORF thereof, or a portion thereof, such as an antigenic region is constructed by in vivo recombination. Methods for in vivo recombination are know in the art and include, for example, the methods disclosed in Chartier, et al. (1996 , J. Virol .
  • Vectors for constructing circovirus genomes include for example, bacterial plasmids which allow multiple copies of the cloned circovirus nucleotide sequence to be produced.
  • the plasmid is co-transfected into a suitable host cell for recombination.
  • suitable host cells for recombination include any cell that will support recombination between a PCV genome and a plasmid containing PCV sequences, or between two or more plasmids, each containing PCV sequences.
  • Recombination is generally performed in procaryotic cells, such as E. coli for example, while generation of circovirus is preferably performed in mammalian cells permissive for PCV replication, such as for example porcine cells and in particular, porcine cells capable of expressing mammalian adenovirus E1 function.
  • the present invention encompasses the use of any mammalian host cell permissive for circovirus replication, and in particular, permissive for replication of PCV, such as PCV1 and PCV2.
  • Allan et al. (1995 , Veterinary Microbiology 44: 49-64) report that PCV replicate in porcine and bovine monocyte/macrophage cultures.
  • Tischer et al. (1987 , Arch. Virol . 96:39-57) report that PCV is known to require actively dividing cells for replication in cell culture.
  • Examples of cells or cell lines useful for replication of PCV include mammalian cells comprising E1 function and permissive for PCV replication, including porcine cells, such as porcine monocyte/macrophage cells and porcine retinal cells, expressing adenovirus E1 function.
  • porcine retina cells expressing human adenovirus E1 function are shown to be permissive for replication of PCV2 and shown to generate virus at 2 ⁇ 10 7 IU/ml.
  • Porcine cell lines are available from public sources such as for example, the American Type Tissue Collection (ATCC). The growth of bacterial cell cultures, as well as culture and maintenance of eukaryotic cells and mammalian cell lines are procedures well-known to those of skill in the art.
  • the present invention encompasses methods of culturing mammalian circovirus, in particular, porcine circovirus, in mammalian host cells transfected with mammalian adenovirus E1 gene sequences.
  • the mammalian cell is stably transformed with adenovirus E1 gene sequences.
  • the E1 gene sequences are integrated into the genome of the mammalian cell.
  • the E1 gene sequences are present on a replicating plasmid.
  • the E1 gene sequence is heterologous to the mammalian cell.
  • a porcine mammalian cell is transformed with a human adenovirus 5 E1 gene sequence.
  • the present invention encompasses the use of any mammalian cell or mammalian cell line expressing E1 function as long as the mammalian cell or cell line expressing E1 function is permissive for the replication of circovirus, in particular porcine circovirus, such as for example, porcine circovirus 1 or porcine circovirus 2.
  • the mammalian cell is a porcine cell or cell line.
  • the present invention encompasses the use of any mammalian E1 function as long as the mammalian host cell expressing the mammalian E1 function is permissive for replication of circovirus, in particular, PCV, such as for example, porcine circovirus 1 or porcine circovirus 2.
  • Mammalian adenovirus genomes are known in the art and are disclosed in, for example, Reddy et al. (1998 , Journal of Virology , 72:1394) which discloses nucleotide sequence, genome organization, and transcription map of bovine adenovirus 3 (BAV3); and Kleiboeker (1995 , Virus Res . 36:259-268), which discloses the E1 region of PAV-4.
  • the present invention encompasses E1 function from any of the various serotypes of human adenovirus, such as Ad2, Ad5, Ad12, and Ad40. In an illustrative embodiment disclosed herein in Example 1, E1 function is human Ad5 E1 function.
  • the human E1A gene is expressed immediately after viral infection (0-2 hours) and before any other viral genes. Flint (1982) Biochem. Biophys. Acta 651:175-208; Flint (1986) Advances Virus Research 31:169-228; Grand (1987) Biochem. J . 241:25-38.
  • the transcription start site of Ad5 E1A is at nucleotide 498 and the ATG start site of the E1A protein is at nucleotide 560 in the virus genome.
  • the E1B protein functions in trans and is necessary for transport of late mRNA from the nucleus to the cytoplasm.
  • the E1B promoter of Ad5 consists of a single high-affinity recognition site for Sp1 and a TATA box.
  • human adenovirus 5 E1A and E1B gene sequences are located at nucleotides 505-4034 of the nucleotide sequence provided in Chroboezek, J. et al. (1992 , Virology . 186:280-285).
  • the mammalian host cell is a porcine host cell transfected with human adenovirus 5 E1 gene sequences.
  • the PCV genome can be isolated from PCV virions, or can comprise a PCV genome that has been inserted into a plasmid, using standard techniques of molecular biology and biotechnology. Cloning of the full-length PCV2 genome into vector pBluescript II KS(+) from Strategene by PCR is described in Liu, et al. (2000 , J. Clin. Microbiol . vol 38:3474-3477). The full-length PCV2 genome DNA can be released from the resulting plasmid upon SacII digestion.
  • circovirus nucleotide sequences into permissive mammalian host cells can be achieved by any method known in the art, including, but not limited to, transfection and transformation including, but not limited to, microinjection, electroporation, CaPO 4 precipitation, DEAE-dextran, liposomes, particle bombardment, etc.
  • transfection and transformation including, but not limited to, microinjection, electroporation, CaPO 4 precipitation, DEAE-dextran, liposomes, particle bombardment, etc.
  • An illustrative method for transfecting PCV2 nucleotide sequences into VIDO R1 cells is described herein in Example 3.
  • procaryotic cells such as bacterial cells
  • eukaryotic cells such as mammalian host cells expressing adenovirus E1 function
  • porcine embryonic retina cells Primary cultures of porcine embryonic retina cells were transfected with 10 ⁇ g of plasmid pTG 4671 (Transgene, France) by the calcium phosphate technique.
  • the pTG 4671 plasmid contains the entire E1A and E1B sequences (nts 505-4034) of HAV-5, along with the puromycin acetyltransferase gene as a selectable marker.
  • the E1 region is under the control of the constitutive promoter from the mouse phosphoglycerate kinase gene, and the puromycin acetyltransferase gene is controlled by the constitutive SV40 early promoter.
  • Transformed cells were selected by three passages in medium containing 7 ⁇ g/ml puromycin, identified based on change in their morphology from single foci (i.e., loss of contact inhibition), and subjected to single cell cloning.
  • the established cell line was first tested for its ability to support the growth of E1 deletion mutants of HAV-5. Subsequently the cell line was further investigated for the presence of E1 sequences in the genome by PCR, expression of the E1A and E1B proteins by Western blot, and doubling time under cell culture conditions. E1 sequences were detected, and production of E1A and E1B proteins was demonstrated by immunoprecipitation. Doubling time was shorter, when compared to that of the parent cell line.
  • VIDO R1 cells were cultured through more than 50 passages (split 1:3 twice weekly) and tested for their ability to support the replication of E1-deleted HAV-5. Expression of the E1A and E1B proteins at regular intervals was also monitored by Western blot. The results indicated that the VIDO R1 line retained the ability to support the growth of E1-deleted virus and expressed similar levels of E1 proteins during more than 50 passages in culture. Therefore, VIDO R1 can be considered to be an established cell line. VIDO R1 cell line has been deposited with the American Type Culture collection (ATCC) and has ATCC accession number PTA-155.
  • ATCC American Type Culture collection
  • Example 2 provides a description of the molecular cloning of full-length PCV2 genome.
  • PCV2 DNA was amplified by PCR from total DNA extracted from a piglet with PMWS.
  • the cloning of the full-length PCV2 genome DNA into vector pBluescript II KS(+) (Stratagene) by polymerase chain reaction (PCR) was described in Liu et al. (2000). J. Clin. Microbiol . 38:3474-3477).
  • the PCV2 sequence was submitted to GenBank (Accession no. AF086834).
  • GenBank accesion no. AF086834
  • the full-length PCV2 genome DNA is released from the resulting plasmid upon SacII digestion.
  • Example 3 describes the transfection of VIDO R1 cells, as described in Example 1, with a plasmid containing the PCV2 genome as constructed in Example 2.
  • Fetal porcine retina cell line, VIDO R1, as described in Example 1 and Vero cells were maintained at 37° C. with 5% CO 2 in Eagles based MEM media supplemented with 10% or 5% heat-inactivated fetal bovine serum (FBS), respectively.
  • FBS heat-inactivated fetal bovine serum
  • Monolayers of VIDO R1 cells grown in a six-well dish were transfected with cloned PCV2 DNA using Lipofectin according to the manufacturer's recommendations (GIBCO BRL). Prior to transfection, PCV2 full-length genome was released from the plasmid by digestion with SacII (Liu, Q., et al., 2000 , J. Clin. Microbiol . 38:3474-3477). For infection, the transfected VIDO R1 cells were subjected to three cycles of freezing ( ⁇ 70° C.) and thawing (37° C.). The lysate was then clarified by centrifugation and used to infect fresh VIDO R1 cells.
  • PCV2-infected VIDO R1 cells were incubated with 0.5% Triton X-114 in phosphate-buffered saline (PBS) at 37° C. for 45 min followed by Freon 113 (1,1,2-trichloro-trifluoroethane) extraction.
  • PBS phosphate-buffered saline
  • Freon 113 (1,1,2-trichloro-trifluoroethane) extraction.
  • the cell debris and membranes were clarified by centrifugation at 2000 g for 15 min.
  • the viruses in the supernatant were pelleted at 35000 g for 3 h through a 20% sucrose cushion.
  • the virus pellet was suspended in PBS and stored at ⁇ 70° C.
  • Virus titers were determined as infectious units (IU) by quantitative ORF2 protein immuno-peroxidase staining.
  • the cell monolayers in 12-well dishes were infected with serial dilutions of virus. After adsorption of virus for 1 h, the cells were washed and overlaid with MEM containing 2% FBS and 0.7% agarose. On day 3 post infection (p.i.), the agarose overlay was removed and the cells were fixed and permeabilized with methanol/acetone (1:1 in volume) for 20 min at ⁇ 20° C. After blocking with 1% bovine serum albumin for 1 h at room temperature, the cells were incubated with rabbit anti-ORF2 serum (Liu et al., 2001, Protein Expression and Purification. 21:115-120).
  • the plates were washed with PBS and then processed using VECTASTAIN Elite ABC kit (Vector Laboratories). The reaction was developed with 3,3′diaminobenzidine (DAB) tetrahydrochloride and observed under a microscope. By counting the positively stained cells, the virus titer was expressed as IU where 1 IU was defined as one positively stained cell/foci at 3 d.p.i.
  • DAB 3,3′diaminobenzidine
  • Viral DNA was extracted from PCV2-infected VIDO R1 cell monolayers by the method of Hirt (1967 , J. Mol. Biol . 26:365-369). The viral DNA was then characterized by restriction analysis and polymerase chain reaction (PCR) as described in Liu et al., (2000). J. Clin. Microbiol . 38:3474-3477).
  • PCR polymerase chain reaction

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
US10/112,540 2001-03-27 2002-03-27 Methods to culture circovirus Expired - Lifetime US6794163B2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US10/112,540 US6794163B2 (en) 2001-03-27 2002-03-27 Methods to culture circovirus
US10/840,879 US7172899B2 (en) 2001-03-27 2004-05-07 Methods to culture circovirus
US11/654,869 US7560278B2 (en) 2001-03-27 2007-01-17 Methods to culture circovirus

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US27917301P 2001-03-27 2001-03-27
US10/112,540 US6794163B2 (en) 2001-03-27 2002-03-27 Methods to culture circovirus

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US10/840,879 Division US7172899B2 (en) 2001-03-27 2004-05-07 Methods to culture circovirus

Publications (2)

Publication Number Publication Date
US20020177216A1 US20020177216A1 (en) 2002-11-28
US6794163B2 true US6794163B2 (en) 2004-09-21

Family

ID=23067939

Family Applications (3)

Application Number Title Priority Date Filing Date
US10/112,540 Expired - Lifetime US6794163B2 (en) 2001-03-27 2002-03-27 Methods to culture circovirus
US10/840,879 Expired - Lifetime US7172899B2 (en) 2001-03-27 2004-05-07 Methods to culture circovirus
US11/654,869 Expired - Lifetime US7560278B2 (en) 2001-03-27 2007-01-17 Methods to culture circovirus

Family Applications After (2)

Application Number Title Priority Date Filing Date
US10/840,879 Expired - Lifetime US7172899B2 (en) 2001-03-27 2004-05-07 Methods to culture circovirus
US11/654,869 Expired - Lifetime US7560278B2 (en) 2001-03-27 2007-01-17 Methods to culture circovirus

Country Status (24)

Country Link
US (3) US6794163B2 (fr)
EP (1) EP1379671B1 (fr)
JP (1) JP4038127B2 (fr)
KR (1) KR100876629B1 (fr)
CN (2) CN1309836C (fr)
AT (1) ATE430807T1 (fr)
AU (1) AU2002244584B2 (fr)
BR (1) BRPI0208456B8 (fr)
CA (1) CA2442135C (fr)
CY (1) CY1109267T1 (fr)
CZ (1) CZ304525B6 (fr)
DE (1) DE60232234D1 (fr)
DK (1) DK1379671T3 (fr)
ES (1) ES2324466T3 (fr)
HU (1) HU228065B1 (fr)
MX (1) MXPA03008810A (fr)
MY (2) MY148759A (fr)
PL (1) PL202951B1 (fr)
PT (1) PT1379671E (fr)
RU (2) RU2312142C2 (fr)
SK (1) SK287969B6 (fr)
UA (1) UA82647C2 (fr)
WO (1) WO2002077210A2 (fr)
ZA (1) ZA200307517B (fr)

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050147966A1 (en) * 2001-06-28 2005-07-07 Virginia Tech Intellectual Properties, Inc. Differential PCR-RFLP assay for detecting and distinguishing between nonpathogenic PCV-1 and pathogenic PCV-2
US20050238662A1 (en) * 1997-12-05 2005-10-27 Agence Francaise De Securite Sanitaire Des Aliments Circovirus sequences associated with piglet weight loss disease (PWD)
US20050239187A1 (en) * 2001-03-27 2005-10-27 Qiang Liu Methods to culture circovirus
US20060222663A1 (en) * 1997-12-05 2006-10-05 Agence Francaise De Securite Sanitaire Des Aliments Circovirus sequences associated with piglet weight loss disease (PWD)
US20080226669A1 (en) * 2005-12-29 2008-09-18 Boehringer Ingelheim Vetmedica, Inc. Multivalent pcv2 immunogenic compositions and methods of producing such compositions
US20080260779A1 (en) * 2005-12-29 2008-10-23 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US20080305128A1 (en) * 2007-01-03 2008-12-11 Boehringer Ingelheim Vetmedica, Inc. Treatment of prdc in pigs
US20090016992A1 (en) * 2004-12-30 2009-01-15 Boehringer Ingelheim Vetmedica, Inc. Pcv2 immunogenic compositions and methods of producing such compositions
US20090098158A1 (en) * 2007-09-04 2009-04-16 Boehringer Ingelheim Vetmedica, Inc. Reduction of concomitant infections in pigs by the use of pcv2 antigen
US20090317423A1 (en) * 2008-01-23 2009-12-24 Boehringer Ingelheim Vetmedica, Inc. Pcv2 mycoplasma hyopneumoniae immunogenic compositions and methods of producing such compositions
US7700285B1 (en) 2005-12-29 2010-04-20 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US20100129397A1 (en) * 2006-12-11 2010-05-27 Boehringer Ingelheim Vetmedica, Inc. Effective method of treatment of porcine circovirus and lawsonia intracellularis infections
US20100167352A1 (en) * 1999-12-03 2010-07-01 Luis Enjuanes Sanchez Infectious clones
US20110008871A1 (en) * 2007-05-11 2011-01-13 Temasek Life Sciences Laboratory Limited Production of Homogeneous Cell Line Highly Permissive To Porcine Circovirus Type 2 (PCV2) Infection
US20110020394A1 (en) * 2007-12-31 2011-01-27 Boehringer Ingelheim Vetmedica, Inc. Pcv2 orf2 virus like particle with foreign amino acid insertion
US20110091499A1 (en) * 2006-12-15 2011-04-21 Boehringer Ingelheim Vetmedica, Inc. Treatment of pigs with pcv2 antigent
US7943298B2 (en) 2007-02-13 2011-05-17 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical PCVD
US20120100168A1 (en) * 2010-01-25 2012-04-26 Blood Systems, Inc. Cyclovirus and methods of use
US9505808B2 (en) 2013-10-02 2016-11-29 Boehringer Ingelheim Vetmedica, Inc. PCV2 ORF2 protein variant and virus like particles composed thereof
US9561270B2 (en) 2009-09-02 2017-02-07 Boehringer Ingelheim Vetmedica, Inc. Methods of reducing virucidal activity in PCV-2 compositions and PCV-2 compositions with an improved immunogenicity
US9919041B2 (en) 2004-12-30 2018-03-20 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US9987348B2 (en) 2013-09-25 2018-06-05 Zoetis Services Llc PCV2B divergent vaccine composition and methods of use

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7276353B2 (en) 2001-12-12 2007-10-02 Virginia Tech Intellectual Properties, Inc. Chimeric infectious DNA clones, chimeric porcine circoviruses and uses thereof
US7279166B2 (en) 2001-12-12 2007-10-09 Virginia Tech Intellectual Properties, Inc. Chimeric infectious DNA clones, chimeric porcine circoviruses and uses thereof
US8834891B2 (en) 2005-03-14 2014-09-16 Boehringer Ingelheim Vetmedica, Inc. Immunogenic compositions comprising Lawsonia intracellularis
WO2008151215A1 (fr) * 2007-06-04 2008-12-11 Merial Limited Production de protéines de circovirus porcin dans des plantes
NZ586238A (en) * 2007-12-21 2012-10-26 Wyeth Llc Methods and compositions for immunizing pigs against porcine circovirus
KR20100135903A (ko) * 2008-04-16 2010-12-27 버지니아 테크 인터렉추얼 프라퍼티스, 인크. 키메라 돼지 써코바이러스 ＰＣＶ２Ｇｅｎ―１Ｒｅｐ 및 이의 용도
US9944904B2 (en) 2015-05-14 2018-04-17 Merial Inc. Method for porcine circovirus production and PCV2 vaccines
WO2018071511A1 (fr) * 2016-10-11 2018-04-19 Orion Labs Transfert de communication de groupe vers un service secondaire
CN114934020B (zh) * 2022-01-30 2024-02-13 山东省滨州畜牧兽医研究院 猪视网膜上皮细胞系及其建立方法和应用

Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999008706A1 (fr) 1997-08-14 1999-02-25 Commonwealth Scientific And Industrial Research Organisation Vecteur d'adenovirus de recombinaison porcin
WO1999018214A1 (fr) 1997-10-03 1999-04-15 Merial Circovirus porcins, vaccins et reactifs de diagnostic
WO1999029871A2 (fr) 1997-12-05 1999-06-17 Agence Francaise De Securite Sanitaire Des Aliments Sequences de circovirus associe a la maladie de l'amaigrissement du porcelet (map)
WO1999045956A1 (fr) 1998-03-13 1999-09-16 University Of Georgia Research Foundation, Inc. Vaccins contre les infections par circovirus
WO1999053047A2 (fr) 1998-04-15 1999-10-21 University Of Saskatchewan Genome d'adenovirus porcin de type 3
WO2000014092A1 (fr) 1998-09-08 2000-03-16 Giri Brij P 1,2-dioxetanes chimioluminescents
WO2000026395A2 (fr) 1998-11-02 2000-05-11 University Of Saskatchewan Cellules bovines exprimant les fonctions essentielles de l'adenovirus pour la propagation de vecteurs recombines de l'adenovirus
WO2000047756A1 (fr) 1999-02-11 2000-08-17 Merial Vecteurs et vaccins viraux a base d'adenovirus porcins recombines
WO2000077216A2 (fr) 1999-06-10 2000-12-21 Merial Vaccin poxvirus recombine contre le circovirus porcin
WO2001016330A2 (fr) 1999-08-31 2001-03-08 Merial Prevention d'une myocardite, d'un avortement et d'une infection intra-uterine associes au circovirus porcin de type 2
US6217883B1 (en) 1998-07-06 2001-04-17 Merial Porcine circovirus and paravovirus vaccine
WO2001083737A2 (fr) 2000-05-03 2001-11-08 University Of Guelph Vaccin d"adenovirus porcin
US6391314B1 (en) 1997-10-03 2002-05-21 Merial Porcine circoviruses vaccines diagnostic reagents

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998017799A1 (fr) * 1996-10-23 1998-04-30 The Trustees Of The University Of Pennsylvania Immunotherapie et vaccins perfectionnes
US7192594B2 (en) * 1997-10-03 2007-03-20 Merial Limited Postweaning multisystemic wasting syndrome and porcine circovirus from pigs
US6943152B1 (en) * 1999-06-10 2005-09-13 Merial DNA vaccine-PCV
WO2002077210A2 (fr) * 2001-03-27 2002-10-03 University Of Saskatchewan Methodes de culture du circovirus

Patent Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999008706A1 (fr) 1997-08-14 1999-02-25 Commonwealth Scientific And Industrial Research Organisation Vecteur d'adenovirus de recombinaison porcin
WO1999018214A1 (fr) 1997-10-03 1999-04-15 Merial Circovirus porcins, vaccins et reactifs de diagnostic
US6368601B1 (en) 1997-10-03 2002-04-09 Merial Porcine circovirus vaccine and diagnostics reagents
US6391314B1 (en) 1997-10-03 2002-05-21 Merial Porcine circoviruses vaccines diagnostic reagents
WO1999029871A2 (fr) 1997-12-05 1999-06-17 Agence Francaise De Securite Sanitaire Des Aliments Sequences de circovirus associe a la maladie de l'amaigrissement du porcelet (map)
WO1999045956A1 (fr) 1998-03-13 1999-09-16 University Of Georgia Research Foundation, Inc. Vaccins contre les infections par circovirus
US6287856B1 (en) 1998-03-13 2001-09-11 University Of Georgia Research Foundation, Inc. Vaccines against circovirus infections
US20030130187A1 (en) 1998-04-15 2003-07-10 Reddy Police Seshidhar Porcine adenovirus type 3 genome
US6492343B1 (en) 1998-04-15 2002-12-10 University Of Saskatchewan Porcine adenovirus type 3 genome
WO1999053047A2 (fr) 1998-04-15 1999-10-21 University Of Saskatchewan Genome d'adenovirus porcin de type 3
US6217883B1 (en) 1998-07-06 2001-04-17 Merial Porcine circovirus and paravovirus vaccine
WO2000014092A1 (fr) 1998-09-08 2000-03-16 Giri Brij P 1,2-dioxetanes chimioluminescents
WO2000026395A2 (fr) 1998-11-02 2000-05-11 University Of Saskatchewan Cellules bovines exprimant les fonctions essentielles de l'adenovirus pour la propagation de vecteurs recombines de l'adenovirus
WO2000047756A1 (fr) 1999-02-11 2000-08-17 Merial Vecteurs et vaccins viraux a base d'adenovirus porcins recombines
WO2000077216A2 (fr) 1999-06-10 2000-12-21 Merial Vaccin poxvirus recombine contre le circovirus porcin
WO2001016330A2 (fr) 1999-08-31 2001-03-08 Merial Prevention d'une myocardite, d'un avortement et d'une infection intra-uterine associes au circovirus porcin de type 2
WO2001083737A2 (fr) 2000-05-03 2001-11-08 University Of Guelph Vaccin d"adenovirus porcin

Non-Patent Citations (54)

* Cited by examiner, † Cited by third party
Title
Allan, G. M. and Ellis, J. A. (2000). "Porcine Circoviruses: A Review," J. Vet. Diagn. Invest. 12:3-14.
Allan, G. M. et al. (1994). "Some Biological and Physico-Chemical Properties of Porcine Circovirus," J. Vet Med. B. 41:17-26.
Allan, G. M. et al. (1995). "Pathogenesis of Porcine Circovirus; Experimental Infections of Colostrum Deprived Piglets and Examination of Pig Foetal Material," Veterinary Microbiology 44:49-64.
Allan, G. M. et al. (1998). "Isolation of Porcine Circovirus-Like Viruses from Pigs with a Wasting Disease in the USA and Europe," in this European J. Vet. Diagn. Invest. 10:3-10.
Chartier, C. et al. (Jul. 1996). "Efficient Generation of Recombinant Adenovirus Vectors by Homologous Recombination in Escherichia Coli," J. Virol. 70(7):4805-4810.
Chroboczek, J. et al. (1992). "The Sequence of the Genome of Adenovirus Type 5 and Its Comparison with the Genome of Adenovirus Type 2," Virology 186:280-285.
Ellis J. et al. (1990). "Reproduction of Lesions of Postweaning Multisystemic Wasting Syndrome in Gnotobiotic Piglets," J. Vet. Diagn. Invest. 11:3-14.
Ellis, J. et al. (Jan. 1998). "Isolation of Circovirus from Lesions of Pigs with Postweaning Multisystemic Wasting Syndrome," Can. Vet. J. 39:44-51.
English translation of PCT publication No. WO/47756, published Aug. 17, 2000, entitled "Recombined Porcine Adenovirus Based Viral Vaccines and Vectors," 60 pages.
English translation of PCT publication No. WO99/29871, entitled Circovirus Sequences Associated With Piglet Weight Loss Disease (PWD), 112 pages.
Flint, S. J. (1986). "Regulation of Adenovirus mRNA Formation," Advances in Virus Research 31:169-228.
Flint, S. J. et al. (1982). "Expression of Adenoviral Genetic Information in Productively Infected Cells," Biochem. Biophys. Acta 651:175-208.
Grand, R. J. A. (1987). "The Structure and Functions of the Adenovirus Early Region 1 Proteins," Biochem. J. 241:25-38.
Hamel, A. L. et al. (Jun. 1998). "Nucleotide Sequence of Porcine Circovirus Associated with Postweaning Multisystemic Wasting Syndrome in Pigs," J. Virol. 72(6):5262-5267.
Hamel, A. L. et al. (May 1998). Genbank Accession No. AF027217. Porcine Circoviru . . . [2689645]. "Nucleotide Sequence of Porcine Circovirus Associated with Postweaning Multisystemic Wasting Syndrome in Pigs,", total pages 4.
Hirt, B. (1967). "Selective Extraction of Polyma DNA from Infected Mouse Cell Cultures," J. Mol. Biol. 26:365-369.
Kennedy, S. et al. (2000). "Reproduction of Lesions of Postweaning Multisystemic Wasting Syndrome by Infection of Conventional Pics with Porcine Circovius Type 2 Alone or in Combination with Porcine Parvovirus," Journal of Comparative Pathology 122(1):9-24.
Kleiboeker, S. B. (1995). "Identification and Sequence Analysis of the E1 Genomic Region of a Porcine Adenovirus," Virus Res. 36:259-268.
Krakowak, S. et al. (2000). "Viral Wasting Syndrome of Swing: Experimental Reproduction of Postweaning Multisystemic Wasting Syndrome in Gnotobiotic Swine by Coinfection with Porcine Circovirus 2 and Porcine Parvovirus," Vet. Pathol. 37:254-263.
Liu, Q et al. (2001). "Bacterial Expression of an Immunologically Reactive PCV2 ORF2 Fusion Protein," Protein Express and Purif. 21:115-120.
Liu, Q et al. (Sep. 2000). "Quantitative Competitive PCR Analysis of Porcine Circovirus DNA in Serum from Pigs with Postweaning Multisystemic Wasting Syndrome," J. Clin. Microbiol. 38(9):3474-3477.
Liu, Q. et al. (2001). "Nuclear Localization of the ORF2 Protein Encoded by Porcine Circovirus Type 2," Virology 285(1):91-99.
Lukert, P. D. et al. (1995). "Circoviridae", In Virus Taxonomy: Classification and Nomenclature of Viruses. F. A. Murphy, et al. (eds.) Sixth Report of the International Committee on Taxonomy of Viruses, Virology Division International Union of Microbiological Societies, Springer-Verlag Wien New York, pp. 166-168.
Mahé, D. et al. (2000). "Differential Recognition of ORF2 Protein from Type 1 and Type 2 Porcine Circoviruses and Identification of Immunorelevant Epitopes," J. Gen. Virol. 81:1815-1824.
Mankertz, A. et al. (1998). "Identification of a Protein Essential for Replication of Porcine Circovirus," J. Gen. Virol. 79:381-384.
Mankertz, A. et al. (2000). "Characterisation of PCV-2 Isolates from Spain, Germany, and France," Virus Res. 66:65-77.
Mankertz, A. et al. (Mar. 1997). "Mapping and Characterization of the Origin of DNA Replication of Porcine Circovirus," J. Virol. 71(3):2562-2566.
Meehan, B. M. et al. (1997). "Sequence of Porcine Circovirus DNA: Affinities with Plant Circoviruses," J. Gen. Virol. 78:221-227.
Meehan, B. M. et al. (1998). "Characterization of Novel Circovirus DNAs Associated with Wasting Syndromes in Pigs," J. Gen. Virol. 79:2171-2179.
Morozov, I. et al. (Sep. 1998). "Detection of a Novel Strain of Porcine Circovirus in Pigs with Postweaning Multisystemic Wasting Syndrome," J. Clin. Microbiol. 36(9):2535-2541.
Murphy, F. A. et al. eds. (1999). "Circoviridae" Chapter 22 In Veterinary Virology. 3rd ed. Academic Press, p. 357-361.
Nagy, M. and Tuboly, T. (2000). "Porcine Adenoviruses: An Update On Genome Analysis and Vector Development," Acta Veterinaria Hungarica 48(4):491-499.
Nagy, M. et al. (2001). "The Complete Nucleotide Sequence of Porcine Adenovirus Serotype 5". Journal of General Virology 82(3):525-529.
Nawagitgul, P. et al. (2000). "Open Reading Frame 2 of Porcine Circovirus Type 2 Encodes a Major Capsid Protein," J. Gen. Virol. 81:2281-2287.
Pogranichnyy, R. M. et al. (2000). "Characterizaton of Immune Response of Young Pigs to Porcine Circovirus Type 2 Infection," Virol. Immunol. 13(2):143-153.
Reddy, P. A. et al. (1999). "Replication-Defective Bovine Adenovirus Type 3 as an Expression Vector," Journal of Virology 73(11):9137-9144.
Reddy, P. S. et al. (1995). "Comparison of the Inverted Terminal Repetition Sequences from Five Porcine Adenovirus Serotypes," Virology 212(1):237-239.
Reddy, P. S. et al. (1999). "Porcine Adenovirus-3 As A Helper-Dependent Expression Vector," J. Gen. Virol. 80:2909-2916.
Reddy, P. S. et al. (Feb. 1998). "Nucleotide Sequence, Genome Organization, and Transcription Map of Bovine Adenovirus Type 3," J. of Virol. 72(2):1394-1402.
Sambrook, J. et al., (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, p. 7.52 entitled "Hybridization and Autoradiography".
Shenk, T. (1996). "Adenoviridae: The Viruses and Their Replications," In Fields Virology. 3rd ed. B.N. Fields, D.M. Knipe and P.M. Howley ed., Lippincott-Raven Publishers, Philadelphia, pp. 2111-2148.
Shenk, T. (1996). "Chapter 30: Adenoviridae: The Viruses and Their Replications," In Fields Virology, 3rd ed. B.N. Fields, D.M. Knipe and P.M. Howley eds., Lippincott-Raven Publishers, Philadephia, pp. 979-1016.
Tischer, I. D. et al. (1995). "Occurrence and Role of an Early Antigen and Evidence for Transforming Ability of Porcine Circovirus," Arch. Virol. 140:1799-1816.
Tischer, I. et al. (1986). "Studies on Epidemiology and Pathogenicity of Porcine Circovirus," Arch. Virol. 91:271-276.
Tischer, I. et al. (1987). "Replication of Porcine Circovirus: Induction by Glucosamine and Cell Cycle Dependence," Arch. Virol. 96:39-57.
Tischer, I. et al. (Jan. 1982). "A Very Small Procine Virus with Circular Single-Stranded DNA," Nature 295:64-66.
Tuboly, T. and Nagy, E. (2000). "Sequence Analysis and Deletion of Porcine Adenovirus Serotype 5 E3 Region," Virus Research 68(2):109-117.
Tuboly, T. et al. (1993). "Potential Viral Vectors for the Stimulation of Mucosal Antibody Responses Against Enteric Viral Antigens in Pigs," Research In Veterniary Science 54(3):345-350.
Tuboly, T. et al. (1995). "Restriction Endonuclease Analysis and Physical Mapping of the Genome of Porcine Adenovirus Type 5," Virus Research 37(1):49-54.
Tuboly, T. et al. (2000). "Characterization of Early Region 4 of Porcine Adenovirus Serotype 5," Virus Genes 20(3): 217-219.
Tuboly, T. et al. (2001). "Construction and Characterization of Recombinant Porcine Adenovirus Serotype 5 Expressing the Transmissible Gastroenteritis Virus Spike Gene," Journal of General Virology 82(1):183-190.
Wang, L. et al. (Sep. 1998). Genbank Accession No. AF086834. Porcine Circovir . . . [gi:3661515]. "Porcine Cirovirus Strain B9, Complete Genome," total pages 4.
Wang, L. et al. (Sep. 1998). Genbank Accession No. AF086835. Porcine Circovir . . . [gi:3661518]. "Porcine Cirovirus Strain 9741, Complete Genome," total pages 3.
Wang, L. et al. (Sep. 1998). Genbank Accession No. AF086836. Porcine Circovir . . . [gi:3661521]. "Porcine Cirovirus Strain M226, Complete Genome," total pages 3.

Cited By (101)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7604808B2 (en) 1997-12-05 2009-10-20 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US20070059324A1 (en) * 1997-12-05 2007-03-15 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US8715690B2 (en) 1997-12-05 2014-05-06 Zoetis W Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US20060222659A1 (en) * 1997-12-05 2006-10-05 Agence Francaise De Securite Sanitaire Des Aliments Circovirus sequences associated with piglet weight loss disease (PWD)
US20060222663A1 (en) * 1997-12-05 2006-10-05 Agence Francaise De Securite Sanitaire Des Aliments Circovirus sequences associated with piglet weight loss disease (PWD)
US20100189736A1 (en) * 1997-12-05 2010-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US20070041990A1 (en) * 1997-12-05 2007-02-22 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US8916353B2 (en) 1997-12-05 2014-12-23 Zoetis W Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US7223407B2 (en) 1997-12-05 2007-05-29 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7758865B2 (en) 1997-12-05 2010-07-20 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7258865B2 (en) 1997-12-05 2007-08-21 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7261898B2 (en) 1997-12-05 2007-08-28 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7740865B2 (en) 1997-12-05 2010-06-22 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7405075B2 (en) 1997-12-05 2008-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7407803B2 (en) 1997-12-05 2008-08-05 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7425444B2 (en) 1997-12-05 2008-09-16 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US10052375B2 (en) 1997-12-05 2018-08-21 Zoetics Services LLC Circovirus sequences associated with piglet weight loss disease (PWD)
US20080233147A1 (en) * 1997-12-05 2008-09-25 Andre Jestin Circovirus sequences associated with piglet weight loss disease (PWD)
US9717784B2 (en) 1997-12-05 2017-08-01 Zoetis Services Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US9700613B2 (en) 1997-12-05 2017-07-11 Zoetis Services Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US7390494B2 (en) 1997-12-05 2008-06-24 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US20050238662A1 (en) * 1997-12-05 2005-10-27 Agence Francaise De Securite Sanitaire Des Aliments Circovirus sequences associated with piglet weight loss disease (PWD)
US7740866B2 (en) 1997-12-05 2010-06-22 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US8415525B2 (en) 1997-12-05 2013-04-09 Zoetis W Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US8124723B2 (en) 1997-12-05 2012-02-28 Wyeth Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US7951907B2 (en) 1997-12-05 2011-05-31 Wyeth Llc Circovirus sequences associated with piglet weight loss disease (PWD)
US20100209453A1 (en) * 1997-12-05 2010-08-19 Wyeth Circovirus sequence associated with piglet weight loss disease (pwd)
US20100203072A1 (en) * 1997-12-05 2010-08-12 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US20090162918A1 (en) * 1997-12-05 2009-06-25 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US7741026B2 (en) 1997-12-05 2010-06-22 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US20100189735A1 (en) * 1997-12-05 2010-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US20100189734A1 (en) * 1997-12-05 2010-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US20100074919A1 (en) * 1997-12-05 2010-03-25 Andre Jestin Circovirus sequences associated with piglet weight loss disease (PWD)
US20100189733A1 (en) * 1997-12-05 2010-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US7722883B2 (en) 1997-12-05 2010-05-25 Wyeth Circovirus sequences associated with piglet weight loss disease (PWD)
US20100189743A1 (en) * 1997-12-05 2010-07-29 Wyeth Circovirus sequences associated with piglet weight loss disease (pwd)
US20100167352A1 (en) * 1999-12-03 2010-07-01 Luis Enjuanes Sanchez Infectious clones
US7560278B2 (en) 2001-03-27 2009-07-14 University Of Saskatchewan Methods to culture circovirus
US20050239187A1 (en) * 2001-03-27 2005-10-27 Qiang Liu Methods to culture circovirus
US20070128700A1 (en) * 2001-03-27 2007-06-07 Qiang Liu Methods to culture circovirus
US7172899B2 (en) 2001-03-27 2007-02-06 University Of Saskatchewan Methods to culture circovirus
US20050147966A1 (en) * 2001-06-28 2005-07-07 Virginia Tech Intellectual Properties, Inc. Differential PCR-RFLP assay for detecting and distinguishing between nonpathogenic PCV-1 and pathogenic PCV-2
US9011872B2 (en) 2004-12-30 2015-04-21 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US7910306B2 (en) 2004-12-30 2011-03-22 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US20090016992A1 (en) * 2004-12-30 2009-01-15 Boehringer Ingelheim Vetmedica, Inc. Pcv2 immunogenic compositions and methods of producing such compositions
US20090022749A1 (en) * 2004-12-30 2009-01-22 Boehringer Ingelheim Vetmedica, Inc. Pcv2 immunogenic compositions and methods of producing such compositions
US10576142B2 (en) 2004-12-30 2020-03-03 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US8025888B2 (en) 2004-12-30 2011-09-27 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US20090022752A1 (en) * 2004-12-30 2009-01-22 Boehringer Ingelheim Vetmedica, Inc. Pcv2 immunogenic compositions and methods of producing such compositions
US7829101B2 (en) 2004-12-30 2010-11-09 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US9919041B2 (en) 2004-12-30 2018-03-20 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US7838213B2 (en) 2005-12-29 2010-11-23 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US20080260779A1 (en) * 2005-12-29 2008-10-23 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US20080226669A1 (en) * 2005-12-29 2008-09-18 Boehringer Ingelheim Vetmedica, Inc. Multivalent pcv2 immunogenic compositions and methods of producing such compositions
US9987349B2 (en) 2005-12-29 2018-06-05 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US9925256B2 (en) 2005-12-29 2018-03-27 Boehringer Ingelheim Vetmedica, Inc. Multivalent PCV2 immunogenic compositions and methods of producing such compositions
US20080279876A1 (en) * 2005-12-29 2008-11-13 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US9925255B2 (en) 2005-12-29 2018-03-27 Boehringer Ingelheim Vetmedica, Inc. Multivalent PCV2 immunogenic compositions and methods of producing such compositions
US20080279875A1 (en) * 2005-12-29 2008-11-13 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US10568955B2 (en) 2005-12-29 2020-02-25 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US7829273B2 (en) 2005-12-29 2010-11-09 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US7968285B2 (en) 2005-12-29 2011-06-28 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US7838214B2 (en) 2005-12-29 2010-11-23 Boehringer Ingelheim Vetmedical, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US20110217327A1 (en) * 2005-12-29 2011-09-08 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US20090092636A1 (en) * 2005-12-29 2009-04-09 Boehringer Ingelheim Vetmedica, Inc Multivalent pcv2 immunogenic compositions and methods of producing such compositions
US8119143B2 (en) 2005-12-29 2012-02-21 Boehringer Ingelheim Vetmedica, Inc. Multivalent PCV2 immunogenic compositions and methods of producing such compositions
US10624963B2 (en) 2005-12-29 2020-04-21 Boehringer Ingelheim Vetmedica, Inc. Multivalent PCV2 immunogenic compositions and methods of producing such compositions
US20080261887A1 (en) * 2005-12-29 2008-10-23 Boehringer Ingelheim Vetmedica, Inc Use of a pcv2 immunogenic composition for lessening clinical symptoms in pigs
US7700285B1 (en) 2005-12-29 2010-04-20 Boehringer Ingelheim Vetmedica, Inc. PCV2 immunogenic compositions and methods of producing such compositions
US9669087B2 (en) 2005-12-29 2017-06-06 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US9011868B2 (en) 2005-12-29 2015-04-21 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US9610345B2 (en) 2005-12-29 2017-04-04 Boehringer Ingelheim Vetmedica, Inc. Use of a PCV2 immunogenic composition for lessening clinical symptoms in pigs
US9101561B2 (en) 2005-12-29 2015-08-11 Boehringer Ingelheim Vetmedica, Inc. Multivalent PCV2 immunogenic compositions and methods of producing such compositions
US9636389B2 (en) 2006-12-11 2017-05-02 Boehringer Ingelheim Vetmedica, Inc. Effective method of treatment of porcine circovirus and Lawsonia intracellularis infections
US20100129397A1 (en) * 2006-12-11 2010-05-27 Boehringer Ingelheim Vetmedica, Inc. Effective method of treatment of porcine circovirus and lawsonia intracellularis infections
US20110091499A1 (en) * 2006-12-15 2011-04-21 Boehringer Ingelheim Vetmedica, Inc. Treatment of pigs with pcv2 antigent
US9517260B2 (en) 2006-12-15 2016-12-13 Boehringer Ingelheim Vetmedica, Inc. Treatment of pigs with PCV2 antigen
US8865183B2 (en) 2006-12-15 2014-10-21 Boehringer Ingelheim Vetmedica, Inc. Treatment of pigs with PCV2 antigent
US20080305128A1 (en) * 2007-01-03 2008-12-11 Boehringer Ingelheim Vetmedica, Inc. Treatment of prdc in pigs
US7914992B2 (en) 2007-01-03 2011-03-29 Boehringer Ingelheim Vetmedica, Inc. Treatment of PRDC in pigs
US9522182B2 (en) 2007-01-03 2016-12-20 Boehringer Ingelheim Vetmedica, Inc. Prophylaxis and treatment of PRDC
US9132187B2 (en) 2007-02-13 2015-09-15 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical PCVD
US7943298B2 (en) 2007-02-13 2011-05-17 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical PCVD
US9555092B2 (en) 2007-02-13 2017-01-31 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical PCVD
US20110182935A1 (en) * 2007-02-13 2011-07-28 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical pcvd
US8496940B2 (en) 2007-02-13 2013-07-30 Boehringer Ingelheim Vetmedica, Inc. Prevention and treatment of sub-clinical PCVD
US20110008871A1 (en) * 2007-05-11 2011-01-13 Temasek Life Sciences Laboratory Limited Production of Homogeneous Cell Line Highly Permissive To Porcine Circovirus Type 2 (PCV2) Infection
US8518696B2 (en) * 2007-05-11 2013-08-27 Temasek Life Sciences Laboratory Limited Production of homogeneous cell line highly permissive to porcine circovirus type 2 (PCV2) infection
US20090098158A1 (en) * 2007-09-04 2009-04-16 Boehringer Ingelheim Vetmedica, Inc. Reduction of concomitant infections in pigs by the use of pcv2 antigen
US8475805B2 (en) 2007-09-04 2013-07-02 Boehringer Ingelheim Vetmedica, Inc. Methods of reducing concomitant infections in pigs with a PCV2 antigen
US7829274B2 (en) 2007-09-04 2010-11-09 Boehringer Ingelheim Vetmedica, Inc. Reduction of concomitant infections in pigs by the use of PCV2 antigen
US20110052629A1 (en) * 2007-09-04 2011-03-03 Boehringer Ingelheim Vetmedica, Inc. Methods of reducing concomitant infections in pigs with a pcv2 antigen
US9132186B2 (en) 2007-09-04 2015-09-15 Boehringer Ingelheim Vetmedica, Inc. Reduction of concomitant infections in pigs by the use of PCV2 antigen
US20110020394A1 (en) * 2007-12-31 2011-01-27 Boehringer Ingelheim Vetmedica, Inc. Pcv2 orf2 virus like particle with foreign amino acid insertion
US9669086B2 (en) 2008-01-23 2017-06-06 Boehringer Ingelheim Vetmedica, Inc. PCV2 Mycoplasma hyopneumoniae immunogenic compositions and methods of producing such compositions
US20090317423A1 (en) * 2008-01-23 2009-12-24 Boehringer Ingelheim Vetmedica, Inc. Pcv2 mycoplasma hyopneumoniae immunogenic compositions and methods of producing such compositions
US9561270B2 (en) 2009-09-02 2017-02-07 Boehringer Ingelheim Vetmedica, Inc. Methods of reducing virucidal activity in PCV-2 compositions and PCV-2 compositions with an improved immunogenicity
US20120100168A1 (en) * 2010-01-25 2012-04-26 Blood Systems, Inc. Cyclovirus and methods of use
US9987348B2 (en) 2013-09-25 2018-06-05 Zoetis Services Llc PCV2B divergent vaccine composition and methods of use
US9505808B2 (en) 2013-10-02 2016-11-29 Boehringer Ingelheim Vetmedica, Inc. PCV2 ORF2 protein variant and virus like particles composed thereof
US11858963B2 (en) 2013-10-02 2024-01-02 Boehringer Ingelheim Animal Health USA Inc. PCV2 ORF2 protein variant and virus like particles composed thereof

Also Published As

Publication number Publication date
WO2002077210A3 (fr) 2003-08-21
ES2324466T3 (es) 2009-08-07
WO2002077210A2 (fr) 2002-10-03
CY1109267T1 (el) 2014-07-02
CA2442135C (fr) 2008-08-19
CN1840656A (zh) 2006-10-04
HU228065B1 (en) 2012-09-28
CN1625598A (zh) 2005-06-08
JP4038127B2 (ja) 2008-01-23
CZ20032584A3 (en) 2004-03-17
KR20040002881A (ko) 2004-01-07
SK287969B6 (sk) 2012-08-06
US20020177216A1 (en) 2002-11-28
UA82647C2 (uk) 2008-05-12
CN1840656B (zh) 2010-06-02
HUP0500161A2 (hu) 2005-05-30
EP1379671B1 (fr) 2009-05-06
RU2007110808A (ru) 2008-09-27
PL364077A1 (en) 2004-12-13
BRPI0208456B1 (pt) 2016-10-18
CZ304525B6 (cs) 2014-06-18
RU2312142C2 (ru) 2007-12-10
MY148759A (en) 2013-05-31
AU2002244584B2 (en) 2006-12-14
US20070128700A1 (en) 2007-06-07
KR100876629B1 (ko) 2009-01-07
RU2003131342A (ru) 2005-04-20
ATE430807T1 (de) 2009-05-15
BRPI0208456B8 (pt) 2021-05-25
DE60232234D1 (de) 2009-06-18
SK12102003A3 (sk) 2004-05-04
DK1379671T3 (da) 2009-06-29
PT1379671E (pt) 2009-07-20
ZA200307517B (en) 2004-09-27
US20050239187A1 (en) 2005-10-27
EP1379671A2 (fr) 2004-01-14
CA2442135A1 (fr) 2002-10-03
PL202951B1 (pl) 2009-08-31
US7560278B2 (en) 2009-07-14
BR0208456A (pt) 2004-03-02
US7172899B2 (en) 2007-02-06
HUP0500161A3 (en) 2010-01-28
CN1309836C (zh) 2007-04-11
MY146518A (en) 2012-08-15
MXPA03008810A (es) 2004-02-17
JP2005500822A (ja) 2005-01-13

Similar Documents

Publication Publication Date Title
US6794163B2 (en) Methods to culture circovirus
AU2002244584A1 (en) Methods to culture circovirus
Liu et al. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2
JP4623964B2 (ja) キメラ感染性ｄｎａクローン、キメラブタサーコウイルスおよびそれらの使用
JP4749547B2 (ja) ブタサーコウイルスおよびパルボウイルスワクチン
JP2012050453A (ja) 組換えａａｖビリオン産生における使用のための補助機能
JP2015109840A (ja) ブタサーコウイルス、ワクチンおよび診断試薬
Pan et al. Development of recombinant porcine parvovirus-like particles as an antigen carrier formed by the hybrid VP2 protein carrying immunoreactive epitope of porcine circovirus type 2
Fan et al. Immunogenicity of empty capsids of porcine circovius type 2 produced in insect cells
EP2155246B1 (fr) Production d'une lignée de cellules homogènes hautement sensibles à une infection par le circovirus porcin de type 2 (pcv2)
Prasetyo et al. Replication of chicken anemia virus (CAV) requires apoptin and is complemented by VP3 of human torque teno virus (TTV)
Shuai et al. Functional exchangeability of the nuclear localization signal (NLS) of capsid protein between PCV1 and PCV2 in vitro: Implications for the role of NLS in viral replication
JP4856694B2 (ja) 組換えａａｖビリオン産生における使用のための補助機能
Weingartl Porcine circovirus structure and replication: a minireview

Legal Events

Date Code Title Description
CC Certificate of correction
AS Assignment

Owner name: UNIVERSITY OF SASKACHEWAN, SASKATCHEWAN

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:LIU, QIANG;TIKOO, SURESH K.;WILLSON, PHILIP;AND OTHERS;REEL/FRAME:013114/0101;SIGNING DATES FROM 20020531 TO 20020613

AS Assignment

Owner name: UNIVERSITY OF SASKATCHEWAN, CANADA

Free format text: CORRECTIVE ASSIGNMENT TO CORRECT THE ASSIGNEES NAME. DOCUMENT PREVIOUSLY RECORDED AT REEL 013114 FRAME 0101;ASSIGNORS:LIU, QIANG;TIKOO, SURESH K.;WILLSON, PHILIP;AND OTHERS;REEL/FRAME:013538/0975;SIGNING DATES FROM 20020531 TO 20020613

STCF Information on status: patent grant

Free format text: PATENTED CASE

FPAY Fee payment

Year of fee payment: 4

FPAY Fee payment

Year of fee payment: 8

FPAY Fee payment

Year of fee payment: 12